#METABOLOMICS WORKBENCH dj2122_20240830_065411 DATATRACK_ID:5157 STUDY_ID:ST003507 ANALYSIS_ID:AN005759 PROJECT_ID:PR002153 VERSION 1 CREATED_ON September 8, 2024, 1:18 am #PROJECT PR:PROJECT_TITLE A NRF2/β3-adrenoreceptor axis drives a sustained antioxidant and metabolic PR:PROJECT_TITLE rewiring through the pentose-phosphate pathway to alleviate cardiac stress PR:PROJECT_SUMMARY Background Cardiac β3-adrenergic receptors (β3AR) are upregulated in diseased PR:PROJECT_SUMMARY hearts and mediate antithetic effects to those of β1AR and β2AR. β3AR PR:PROJECT_SUMMARY agonists were recently shown to protect from myocardial remodeling in PR:PROJECT_SUMMARY preclinical studies and to improve systolic function in patients with severe PR:PROJECT_SUMMARY heart failure. The underlying mechanisms, however, remain elusive. Methods To PR:PROJECT_SUMMARY dissect functional, transcriptional and metabolic effects, hearts and isolated PR:PROJECT_SUMMARY ventricular myocytes from mice harboring a moderate, cardiac-specific expression PR:PROJECT_SUMMARY of a human ADRB3 transgene (β3AR-Tg) and subjected to transverse aortic PR:PROJECT_SUMMARY constriction (TAC) were assessed using echocardiography, RNAseq, PET scan, PR:PROJECT_SUMMARY metabolomics, seahorse and metabolic flux analysis. Subsequently, signaling and PR:PROJECT_SUMMARY metabolic pathways were investigated further in vivo in β3AR-Tg and in vitro in PR:PROJECT_SUMMARY neonatal rat ventricular myocytes adenovirally infected to express β3AR and PR:PROJECT_SUMMARY subjected to neurohormonal stress. These results were completed with an analysis PR:PROJECT_SUMMARY of single nucleus RNAseq data from human cardiac myocytes from heart failure PR:PROJECT_SUMMARY patients. Results Compared with WT littermate, β3AR-Tg mice were protected from PR:PROJECT_SUMMARY hypertrophy after transaortic constriction (TAC), while systolic function was PR:PROJECT_SUMMARY preserved. β3AR-expressing hearts displayed enhanced myocardial glucose uptake PR:PROJECT_SUMMARY under stress in absence of increased lactate levels. Instead, metabolomic and PR:PROJECT_SUMMARY metabolic flux analyses in stressed hearts revealed an increase in intermediates PR:PROJECT_SUMMARY of the Pentose-Phosphate Pathway (PPP) in β3AR-Tg, an alternative route of PR:PROJECT_SUMMARY glucose utilization, paralleled with increased transcript levels of PR:PROJECT_SUMMARY NADPH-producing and rate-limiting enzymes of the PPP, without fueling the PR:PROJECT_SUMMARY hexosamine metabolism. The ensuing increased content of NADPH and of reduced PR:PROJECT_SUMMARY glutathione decreased myocyte oxidant stress, while downstream oxidative PR:PROJECT_SUMMARY metabolism assessed by oxygen consumption was preserved with higher glucose PR:PROJECT_SUMMARY oxidation in β3AR-Tg post-TAC compared to WT, together with increased PR:PROJECT_SUMMARY mitochondrial biogenesis. Unbiased transcriptomics and pathway analysis PR:PROJECT_SUMMARY identified NRF2 (NFE2L2) as upstream transcription factor which was functionally PR:PROJECT_SUMMARY verified in β3AR-expressing cardiac myocytes where its translocation and PR:PROJECT_SUMMARY nuclear activity was dependent on β3AR activation of nitric-oxide synthase PR:PROJECT_SUMMARY (NOS) NO production. Conclusion Moderate expression of cardiac β3AR, at levels PR:PROJECT_SUMMARY observed in human cardiac myocardium, exerts antioxidant effects through PR:PROJECT_SUMMARY activation of the PPP and NRF2 pathway, thereby preserving myocardial oxidative PR:PROJECT_SUMMARY metabolism, function and integrity under pathophysiological stress. PR:INSTITUTE UCLouvain PR:LAST_NAME Dewulf PR:FIRST_NAME Joseph PR:ADDRESS Avenue Hippocrate 10, Brussels, Brussels, 1200, Belgium PR:EMAIL joseph.dewulf@uclouvain.be PR:PHONE 027646727 #STUDY ST:STUDY_TITLE A NRF2/β3-adrenoreceptor axis drives a sustained antioxidant and metabolic ST:STUDY_TITLE rewiring through the pentose-phosphate pathway to alleviate cardiac stress ST:STUDY_SUMMARY WT TAC vs TG TAC vs WT baseline vs TG baseline Background: Cardiac ST:STUDY_SUMMARY β3-adrenergic receptors (β3AR) are upregulated in diseased hearts and mediate ST:STUDY_SUMMARY antithetic effects to those of β1AR and β2AR. β3AR agonists were recently ST:STUDY_SUMMARY shown to protect from myocardial remodeling in preclinical studies and to ST:STUDY_SUMMARY improve systolic function in patients with severe heart failure. The underlying ST:STUDY_SUMMARY mechanisms, however, remain elusive. Methods: To dissect functional, ST:STUDY_SUMMARY transcriptional and metabolic effects, hearts and isolated ventricular myocytes ST:STUDY_SUMMARY from mice harboring a moderate, cardiac-specific expression of a human ADRB3 ST:STUDY_SUMMARY transgene (β3AR-Tg) and subjected to transverse aortic constriction (TAC) were ST:STUDY_SUMMARY assessed using echocardiography, RNAseq, PET scan, metabolomics, seahorse and ST:STUDY_SUMMARY metabolic flux analysis. Subsequently, signaling and metabolic pathways were ST:STUDY_SUMMARY investigated further in vivo in β3AR-Tg and in vitro in neonatal rat ST:STUDY_SUMMARY ventricular myocytes adenovirally infected to express β3AR and subjected to ST:STUDY_SUMMARY neurohormonal stress. These results were completed with an analysis of single ST:STUDY_SUMMARY nucleus RNAseq data from human cardiac myocytes from heart failure patients. ST:STUDY_SUMMARY Results: Compared with WT littermate, β3AR-Tg mice were protected from ST:STUDY_SUMMARY hypertrophy after transaortic constriction (TAC), while systolic function was ST:STUDY_SUMMARY preserved. β3AR-expressing hearts displayed enhanced myocardial glucose uptake ST:STUDY_SUMMARY under stress in absence of increased lactate levels. Instead, metabolomic and ST:STUDY_SUMMARY metabolic flux analyses in stressed hearts revealed an increase in intermediates ST:STUDY_SUMMARY of the Pentose-Phosphate Pathway (PPP) in β3AR-Tg, an alternative route of ST:STUDY_SUMMARY glucose utilization, paralleled with increased transcript levels of ST:STUDY_SUMMARY NADPH-producing and rate-limiting enzymes of the PPP, without fueling the ST:STUDY_SUMMARY hexosamine metabolism. The ensuing increased content of NADPH and of reduced ST:STUDY_SUMMARY glutathione decreased myocyte oxidant stress, while downstream oxidative ST:STUDY_SUMMARY metabolism assessed by oxygen consumption was preserved with higher glucose ST:STUDY_SUMMARY oxidation in β3ARTg post-TAC compared to WT, together with increased ST:STUDY_SUMMARY mitochondrial biogenesis. Unbiased transcriptomics and pathway analysis ST:STUDY_SUMMARY identified NRF2 (NFE2L2) as upstream transcription factor which was functionally ST:STUDY_SUMMARY verified in β3AR- expressing cardiac myocytes where its translocation and ST:STUDY_SUMMARY nuclear activity was dependent on β3AR activation of nitric-oxide synthase ST:STUDY_SUMMARY (NOS) NO production. Conclusion: Moderate expression of cardiac β3AR, at levels ST:STUDY_SUMMARY observed in human cardiac myocardium, exerts antioxidant effects through ST:STUDY_SUMMARY activation of the PPP and NRF2 pathway, thereby preserving myocardial oxidative ST:STUDY_SUMMARY metabolism, function and integrity under pathophysiological stress. ST:INSTITUTE UCLouvain ST:LAST_NAME Dewulf ST:FIRST_NAME Joseph ST:ADDRESS Avenue Hippocrate 10 ST:EMAIL joseph.dewulf@uclouvain.be ST:PHONE 027646727 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 026 sample 14 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=026 sample 14 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 027 sample 15 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=027 sample 15 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 028 sample 16 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=028 sample 16 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 029 sample 17 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=029 sample 17 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 030 sample 18 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=030 sample 18 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 031 sample 19 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=031 sample 19 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 032 sample 20 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=032 sample 20 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 033 sample 21 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=033 sample 21 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 034 sample 22 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=034 sample 22 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 035 sample 23 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=035 sample 23 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 036 sample 24 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=036 sample 24 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 037 sample 25 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=037 sample 25 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 038 sample 26 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=038 sample 26 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 039 sample 27 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=039 sample 27 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 040 sample 28 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=040 sample 28 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 041 sample 29 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=041 sample 29 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 042 sample 30 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=042 sample 30 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 043 sample 31 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=043 sample 31 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 044 sample 32 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=044 sample 32 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 013 sample 1 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=013 sample 1 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 014 sample 2 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=014 sample 2 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 015 sample 3 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=015 sample 3 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 016 sample 4 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=016 sample 4 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 019 sample 7 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=019 sample 7 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 020 sample 8 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=020 sample 8 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 021 sample 9 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=021 sample 9 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 022 sample 10 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=022 sample 10 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 023 sample 11 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=023 sample 11 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 024 sample 12 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=024 sample 12 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 025 sample 13 PREM M2_ MSE_res_pos IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=025 sample 13 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 026 sample 14 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=026 sample 14 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 027 sample 15 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=027 sample 15 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 028 sample 16 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=028 sample 16 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 029 sample 17 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=029 sample 17 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 030 sample 18 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=030 sample 18 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 031 sample 19 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=031 sample 19 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 032 sample 20 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=032 sample 20 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 033 sample 21 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=033 sample 21 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 034 sample 22 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=034 sample 22 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 035 sample 23 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=035 sample 23 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 036 sample 24 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=036 sample 24 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 037 sample 25 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=037 sample 25 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 038 sample 26 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=038 sample 26 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 039 sample 27 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=039 sample 27 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 040 sample 28 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=040 sample 28 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 041 sample 29 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT TAC 9W RAW_FILE_NAME(Raw file name)=041 sample 29 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 042 sample 30 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=042 sample 30 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 043 sample 31 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=043 sample 31 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 044 sample 32 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG TAC 9W RAW_FILE_NAME(Raw file name)=044 sample 32 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 013 sample 1 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=013 sample 1 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 014 sample 2 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=014 sample 2 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 015 sample 3 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=015 sample 3 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 016 sample 4 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=016 sample 4 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 019 sample 7 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=019 sample 7 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 020 sample 8 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=020 sample 8 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 021 sample 9 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=021 sample 9 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 022 sample 10 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=022 sample 10 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 023 sample 11 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:TG Baseline RAW_FILE_NAME(Raw file name)=023 sample 11 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 024 sample 12 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=024 sample 12 M2_ MSE_res_neg IDC10_ 3ul inj.raw SUBJECT_SAMPLE_FACTORS - 025 sample 13 M2_ MSE_res_neg IDC10_ 3ul inj Sample source:mouse serum | Condition:WT Baseline RAW_FILE_NAME(Raw file name)=025 sample 13 M2_ MSE_res_neg IDC10_ 3ul inj.raw #COLLECTION CO:COLLECTION_SUMMARY Metabolites from serum samples were extracted in microcentrifuge tubes after the CO:COLLECTION_SUMMARY addition of 3 volumes of 100% acetonitrile containing 3 internal standards CO:COLLECTION_SUMMARY [caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic CO:COLLECTION_SUMMARY acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed, CO:COLLECTION_SUMMARY sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10 CO:COLLECTION_SUMMARY minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube CO:COLLECTION_SUMMARY and 3 volumes of pure acetonitrile were added to the initial pellet for a second CO:COLLECTION_SUMMARY similar extraction. The combined upper-phases resulting from the two extractions CO:COLLECTION_SUMMARY were mixed and divided into four equal parts. Each part was dried down under a CO:COLLECTION_SUMMARY gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to CO:COLLECTION_SUMMARY analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v), CO:COLLECTION_SUMMARY 0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75 CO:COLLECTION_SUMMARY (v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the CO:COLLECTION_SUMMARY method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”, CO:COLLECTION_SUMMARY respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to CO:COLLECTION_SUMMARY remove any particulates before being transferred into polypropylene autosampler CO:COLLECTION_SUMMARY vials. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Metabolites from serum samples were extracted in microcentrifuge tubes after the TR:TREATMENT_SUMMARY addition of 3 volumes of 100% acetonitrile containing 3 internal standards TR:TREATMENT_SUMMARY [caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic TR:TREATMENT_SUMMARY acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed, TR:TREATMENT_SUMMARY sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10 TR:TREATMENT_SUMMARY minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube TR:TREATMENT_SUMMARY and 3 volumes of pure acetonitrile were added to the initial pellet for a second TR:TREATMENT_SUMMARY similar extraction. The combined upper-phases resulting from the two extractions TR:TREATMENT_SUMMARY were mixed and divided into four equal parts. Each part was dried down under a TR:TREATMENT_SUMMARY gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to TR:TREATMENT_SUMMARY analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v), TR:TREATMENT_SUMMARY 0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75 TR:TREATMENT_SUMMARY (v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the TR:TREATMENT_SUMMARY method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”, TR:TREATMENT_SUMMARY respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to TR:TREATMENT_SUMMARY remove any particulates before being transferred into polypropylene autosampler TR:TREATMENT_SUMMARY vials. L #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites from serum samples were extracted in microcentrifuge tubes after the SP:SAMPLEPREP_SUMMARY addition of 3 volumes of 100% acetonitrile containing 3 internal standards SP:SAMPLEPREP_SUMMARY [caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic SP:SAMPLEPREP_SUMMARY acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed, SP:SAMPLEPREP_SUMMARY sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10 SP:SAMPLEPREP_SUMMARY minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube SP:SAMPLEPREP_SUMMARY and 3 volumes of pure acetonitrile were added to the initial pellet for a second SP:SAMPLEPREP_SUMMARY similar extraction. The combined upper-phases resulting from the two extractions SP:SAMPLEPREP_SUMMARY were mixed and divided into four equal parts. Each part was dried down under a SP:SAMPLEPREP_SUMMARY gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to SP:SAMPLEPREP_SUMMARY analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v), SP:SAMPLEPREP_SUMMARY 0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75 SP:SAMPLEPREP_SUMMARY (v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the SP:SAMPLEPREP_SUMMARY method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”, SP:SAMPLEPREP_SUMMARY respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to SP:SAMPLEPREP_SUMMARY remove any particulates before being transferred into polypropylene autosampler SP:SAMPLEPREP_SUMMARY vials. L #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY M1POS CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Waters Acquity Premier CH:COLUMN_NAME Waters Acquity Premier BEH amide 1.7um, 2.1 x 100 mm CH:SOLVENT_A water: acetonitrile (93:7), 10mM Ammonium formate pH3 CH:SOLVENT_B water: acetonitrile (7:93), 10mM Ammonium formate pH3 CH:FLOW_GRADIENT 0.2 min 0%A, 20% over 8.3 min, 40% over 1min CH:FLOW_RATE 0.7 ml/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Synapt-XS MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Acquisition: Masslynx The separation columns, operated at 40°C, were an Acquity MS:MS_COMMENTS Premier HSS T3 column 1,8 µm, 2,1 x 100 mm (Waters) and an Acquity Premier BEH MS:MS_COMMENTS Amide 1,8 µm, 2,1 x 100 mm (Waters), for reverse phase and HILIC MS:MS_COMMENTS chromatography, respectively. Mass spectrometry data was acquired in profile MS:MS_COMMENTS (continuum) format over the mass range of m/z 50-1200 using a Waters Synapt XS MS:MS_COMMENTS high resolution Q-TOF mass spectrometer set in MSE resolution mode (scan time MS:MS_COMMENTS 0,1 sec, the low and high trap collision energy were 4V and a ramp between 20 MS:MS_COMMENTS and 50 V, respectively). A dual electrospray ionization (ESI) source was used in MS:MS_COMMENTS positive or negative mode. Capillary and sampling cone, voltages were set to 1 MS:MS_COMMENTS kV and 30 V or 1 kV and 25 V in positive and negative mode, respectively. Source MS:MS_COMMENTS temperature was set to 150 °C and desolvation temperature to 600 °C (550°C in MS:MS_COMMENTS negative ESI). Gas flow rates were set at 1200 L/h for the desolvation gas (1100 MS:MS_COMMENTS L/h in negative ESI) and 50 L/h for the cone gas. Acquisition of leucine MS:MS_COMMENTS enkephalin infused at 10ul/min through a lockspray probe allowed a real time MS:MS_COMMENTS mass correction (30 sec scan intervals). Data processing: Raw LC-MS data were MS:MS_COMMENTS imported and processed in Progenesis QI software (Nonlinear Dynamics), which MS:MS_COMMENTS performed chromatogram alignment, peak picking, ion deconvolution, MS:MS_COMMENTS normalization, peak annotation and statistical analysis. Thresholds of area MS:MS_COMMENTS under the curve > 1000 were set and yielded the following number of detected MS:MS_COMMENTS metabolites: for Control WT vs Control TG (M1 neg : 1886 ; M1 pos : 3881 ; M2 MS:MS_COMMENTS neg : 785 ; M2 pos : 2411) TAC 9W WT vs Control TG (M1 neg : 2022 ; M1 pos : MS:MS_COMMENTS 3690 ; M2 neg : 905 ; M2 pos : 2405) and Control WT vs TAC WT (M1 neg : 2165 ; MS:MS_COMMENTS M1 pos : 4068 ; M2 neg : 944 ; M2 pos : 2448). Volcano plot and FDR threshold of MS:MS_COMMENTS significance were performed on Graphpad Prism. MS:MS_RESULTS_FILE ST003507_AN005759_Results.txt UNITS:arbitrary unit Has m/z:Yes Has RT:Yes RT units:Minutes #END